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      Assessing baloxavir susceptibility of influenza viruses circulating in the United States during the 2016/17 and 2017/18 seasons

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          Abstract

          The anti-influenza therapeutic baloxavir targets cap-dependent endonuclease activity of polymerase acidic (PA) protein. We monitored baloxavir susceptibility in the United States with next generation sequencing analysis supplemented by phenotypic one-cycle infection assay. Analysis of PA sequences of 6,891 influenza A and B viruses collected during 2016/17 and 2017/18 seasons showed amino acid substitutions: I38L (two A(H1N1)pdm09 viruses), E23G (two A(H1N1)pdm09 viruses) and I38M (one A(H3N2) virus); conferring 4–10-fold reduced susceptibility to baloxavir.

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          Baloxavir: First Global Approval

          Young-A Heo (2018)
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            Susceptibility of Influenza Viruses to the Novel Cap-Dependent Endonuclease Inhibitor Baloxavir Marboxil

            Emi Takashita, Hiroko Morita, Rie Ogawa … (2018)
            The novel cap-dependent endonuclease inhibitor baloxavir marboxil was approved for the treatment of influenza A and B virus infections in February 2018 in Japan. Because of the need to monitor influenza viruses for reduced susceptibility to this drug, we used two cell-based screening systems – a conventional plaque reduction assay and a focus reduction assay – to evaluate the susceptibility of influenza viruses to baloxavir. First, we generated a reference virus possessing an I38T substitution in the polymerase acidic subunit (PA), which is known to be associated with reduced susceptibility to baloxavir, and demonstrated the validity of our systems using this reference virus. We then determined the susceptibility of a panel of neuraminidase (NA) inhibitor-resistant viruses and their sensitive counterparts to baloxavir. No significant differences in baloxavir susceptibilities were found between the NA inhibitor-resistant and -sensitive viruses. We also examined seasonal influenza viruses isolated during the 2017–2018 influenza season in Japan and found that no currently circulating A(H1N1)pdm09, A(H3N2), or B viruses had significantly reduced susceptibility to baloxavir and none of the viruses possessed an amino acid substitution at PA residue 38. Use of a combination of methods to analyze antiviral susceptibility and detect amino acid substitutions is valuable for monitoring the emergence of baloxavir-resistant viruses.
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              Insights into the antigenic advancement of influenza A(H3N2) viruses, 2011–2018

              Patricia Jorquera, Vasiliy Mishin, Anton P. Chesnokov … (2019)
              Influenza A(H3N2) viruses evade human immunity primarily by acquiring antigenic changes in the haemagglutinin (HA). HA receptor-binding features of contemporary A(H3N2) viruses hinder traditional antigenic characterization using haemagglutination inhibition and promote selection of HA mutants. Thus, alternative approaches are needed to reliably assess antigenic relatedness between circulating viruses and vaccines. We developed a high content imaging-based neutralization test (HINT) to reduce antigenic mischaracterization resulting from virus adaptation to cell culture. Ferret reference antisera were raised using clinical specimens containing viruses representing recent vaccine strains. Analysis of viruses circulating during 2011–2018 showed that gain of an N158-linked glycosylation in HA was a molecular determinant of antigenic distancing between A/Hong Kong/4801/2014-like (clade 3C.2a) and A/Texas/50/2012-like viruses (clade 3C.1), while multiple evolutionary HA F193S substitution were linked to antigenic distancing from A/Switzerland/97152963/2013-like (clade 3C.3a) and further antigenic distancing from A/Texas/50/2012-like viruses. Additionally, a few viruses carrying HA T135K and/or I192T showed reduced neutralization by A/Hong Kong/4801/2014-like antiserum. Notably, this technique elucidated the antigenic characteristics of clinical specimens, enabling direct characterization of viruses produced in vivo, and eliminating in vitro culture, which rapidly alters the genotype/phenotype. HINT is a valuable new antigenic analysis tool for vaccine strain selection.
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                Author and article information

                Journal
                Journal ID (nlm-ta): Euro Surveill
                Journal ID (iso-abbrev): Euro Surveill
                Journal ID (publisher-id): ES
                Title: Eurosurveillance
                Publisher: European Centre for Disease Prevention and Control (ECDC)
                ISSN (Print): 1025-496X
                ISSN (Electronic): 1560-7917
                Publication date (Print): 17 January 2019
                Volume: 24
                Issue: 3
                Electronic Location Identifier: 1800666
                Affiliations
                [1 ]Influenza Division, National Center for Immunization and Respiratory Diseases, Centers for Disease Control and Prevention (CDC), Atlanta, United States of America
                [2 ]Battelle Memorial Institute, Atlanta, United States of America
                [3 ]Illinois Department of Public Health, Springfield, United States of America
                Author notes

                Correspondence: Larisa V Gubareva ( lgubareva@ 123456cdc.gov )

                Author information
                http://orcid.org/0000-0002-3136-5550
                Article
                Publisher ID: 1800666 Publisher ID: 1800666
                DOI: 10.2807/1560-7917.ES.2019.24.3.1800666
                PMC ID: 6344838
                PubMed ID: 30670144
                SO-VID: 52283143-7829-4102-a3ea-9fff35d4127f
                Copyright © This article is copyright of the authors or their affiliated institutions, 2019.
                License:

                This is an open-access article distributed under the terms of the Creative Commons Attribution (CC BY 4.0) Licence. You may share and adapt the material, but must give appropriate credit to the source, provide a link to the licence, and indicate if changes were made.

                History
                Date received : 12 December 2018
                Date accepted : 16 January 2019
                Categories
                Subject: Rapid Communication

                Keywords: cap-dependent endonuclease inhibitor,polymerase acidic protein,influenza surveillance,influenza,drug resistance,phenotypic testing,next generation sequencing
                Data availability:
                Keywords: cap-dependent endonuclease inhibitor, polymerase acidic protein, influenza surveillance, influenza, drug resistance, phenotypic testing, next generation sequencing

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